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Contents 1 Miniprep for yneA, pGFPrrnB and pSB1C3 1.1 Aim 1.2 Materials and Protocol 1.3 Results 1.3.1 Discussion 1.3.2 Conclusion 2 Transformation of ligated yneA into pGFPrrnB and pSB1C3 2.1 Aims 2.2 Materials and Protocol 3 Plating Bacillus subtilis BFS678

Miniprep for yneA, pGFPrrnB and pSB1C3

Aim

The aim of this experiment is to produce stocks of yneA, pGFPrrnB and pSB1C3.

Materials and Protocol

Please refer to miniprep and nanodrop.

Results

Sample no.	yneA	pGFPrrnB	pSB1C3
1	432.5	238.9	126.1
2	347.6	230.7	107.6
3	380.2	390.9	121.5
4	377.9	236.2	112.8

Table 1: Nanodrop spectrophotometer results. Table represents the amount of plasmid present in µl/ml quantity.

Discussion

Results from the NanoDrop show concentration values of more than 100 ng/µl, which means we have obtained good concentration of DNA from the miniprep.

Conclusion

We keep the miniprep products as stocks for future use.

Transformation of ligated yneA into pGFPrrnB and pSB1C3

Aims

To produce more colonies of the plasmid pGFPrrnB and pSB1C3 containing yneA with pGFPrrnB and pSB1C3 fromyesterday.

Materials and Protocol

Please refer to transformation of E. coli.

Plating Bacillus subtilis BFS678

Bacillus subtilis BFS678 has the pMutin4 (and thus lacI) integrated into the chromosome which will be useful for characterisation. Therefore we will be using this strain for transformation with our filamentous cell and arginase BioBricks.